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      Recombination and Coronavirus Defective Interfering RNAs

      research-article
      a , b
      Seminars in Virology
      Academic Press.
      RNA recombination, leader fusion, recombinant coronaviruses

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          Abstract

          Naturally occurring defective interfering RNAs have been found in 4 of 14 coronavirus species. They range in size from 2.2 kb to approximately 25 kb, or 80% of the 30-kb parent virus genome. The large DI RNAs do not in all cases appear to require helper virus for intracellular replication and it has been postulated that they may on their own function as agents of disease. Coronavirus DI RNAs appear to arise by internal deletions (through nonhomologous recombination events) on the virus genome or on DI RNAs of larger size by a polymerase strand-switching (copy-choice) mechanism. In addition to their use in the study of virus RNA replication and virus assembly, coronavirus DI RNAs are being used in a major way to study the mechanism of a high-frequency, site-specific RNA recombination event that leads to leader acquisition during virus replication (i.e., the leader fusion event that occurs during synthesis of subgenomic mRNAs, and the leader-switching event that can occur during DI RNA replication), a distinguishing feature of coronaviruses (and arteriviruses). Coronavirus DI RNAs are also being engineered as vehicles for the generation of targeted recombinants of the parent virus genome.

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          Most cited references69

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          Coronaviruses: structure and genome expression.

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            RNA recombination in animal and plant viruses.

            Tsz M. Lai (1992)
            An increasing number of animal and plant viruses have been shown to undergo RNA-RNA recombination, which is defined as the exchange of genetic information between nonsegmented RNAs. Only some of these viruses have been shown to undergo recombination in experimental infection of tissue culture, animals, and plants. However, a survey of viral RNA structure and sequences suggests that many RNA viruses were derived form homologous or nonhomologous recombination between viruses or between viruses and cellular genes during natural viral evolution. The high frequency and widespread nature of RNA recombination indicate that this phenomenon plays a more significant role in the biology of RNA viruses than was previously recognized. Three types of RNA recombination are defined: homologous recombination; aberrant homologous recombination, which results in sequence duplication, insertion, or deletion during recombination; and nonhomologous (illegitimate) recombination, which does not involve sequence homology. RNA recombination has been shown to occur by a copy choice mechanism in some viruses. A model for this recombination mechanism is presented.
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              High-frequency RNA recombination of murine coronaviruses.

              The RNA genome of coronaviruses consists of a single species of nonsegmented RNA. In this communication, we demonstrate that the RNA genomes of different strains of murine coronaviruses recombine during mixed infection at a very high frequency. Susceptible cells were coinfected with a temperature-sensitive mutant of one strain of mouse hepatitis virus (MHV) and a wild-type virus of a different strain. Of 21 randomly isolated viruses released from the coinfected cells at the nonpermissive temperature, 2 were recombinants which differed in the site of recombination. After three serial passages of the original virus pool derived from the mixed infection, the majority of the progeny viruses were recombinants. These recombinant viruses represented at least five different recombination sites between the two parental MHV strains. Such a high-frequency recombination between nonsegmented RNA genomes of MHV suggests that segmented RNA intermediates might be generated during MHV replication. We propose that the RNA replication of MHV proceeds in a discontinuous and nonprocessive manner, thus generating free segmented RNA intermediates, which could be used in RNA recombination via a copy-choice mechanism.
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                Author and article information

                Journal
                Semin. Virol
                Seminars in Virology
                Academic Press.
                1044-5773
                1098-1292
                25 May 2002
                1997
                25 May 2002
                : 8
                : 2
                : 101-111
                Affiliations
                [a ]Department of Microbiology, College of Veterinary Medicine, M409 Walters Life Sciences Building, University of Tennessee, Knoxville, Tennessee, 37996-0845
                [b ]Department of Virology, Institute of Medical Microbiology, Leiden University, 2300, RC Leiden, The Netherlands
                Article
                S1044-5773(97)90109-8
                10.1006/smvy.1997.0109
                7129747
                32288442
                b43a250f-6323-4e1b-bd93-9206409d0b9d
                Copyright © 1997 Academic Press. All rights reserved.

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                rna recombination,leader fusion,recombinant coronaviruses

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